Isabelle Benoit, Michèle Asther, Yves Bourne, David Navarro, Stéphane Canaan, Laurence Lesage-Meessen, Marga Herweijer, Pedro M Coutinho, Marcel Asther, Eric Record
The full-length gene that encodes the chlorogenic acid hydrolase from Aspergillus niger CIRM BRFM 131 was cloned by PCR based on the genome of the strain A. niger CBS 513.88. The complete gene consists of 1,715 bp and codes for a deduced protein of 512 amino acids with a molecular mass of 55,264 Da and an acidic pI of 4.6. The gene was successfully cloned and overexpressed in A. niger to yield 1.25 g liter(-1), i.e., 330-fold higher than the production of wild-type strain A. niger CIRM BRFM131. The histidine-tagged recombinant ChlE protein was purified to homogeneity via a single chromatography step, and its main biochemical properties were characterized...
September 2007: Applied and Environmental Microbiology